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Lipooligosaccharides are glycolipids found in the cell wall of many mycobacterial species including the opportunistic pathogen  Mycobacterium kansasii . The genome of  M .  kansasii  ATCC12478 contains a cluster with genes orthologous to  Mycobacterium marinum  LOS biosynthesis genes. To initiate a genetic dissection of this cluster and demonstrate its role in LOS biosynthesis in  M .  kansasii , we chose  MKAN27435 , a gene encoding a putative glycosyltransferase. Using Specialized Transduction, a phage-based gene knockout tool previously used to generate null mutants in other mycobacteria, we generated a  MKAN27435  null mutant. The mutant strain was found to be defective in the biosynthesis of higher LOS subspecies,  viz  LOS-IV, LOS-V, LOS-VI and LOS-VII. Additionally, a range of low abundance species were detected in the mutant strain and mass spectroscopic analysis indicated that these were shunt products generated from LOS-III by the addition of up to six molecules of a pentose.

MKAN27435 Is Required for the Biosynthesis of Higher Subclasses of Lipooligosaccharides in Mycobacterium kansasii